Application of a quantum cascade laser-based spectrometer in a closed chamber system for real-time δC and δO measurements of soil-respired CO2

Laser spectroscopy is an emerging technique to analyze the stable isotopic composition of soil-respired CO₂ (δ¹³C_resp, δ¹⁸O_resp) in situ and at high temporal resolution. Here we present the first application of a quantum cascade laser-based spectrometer (QCLS) in a closed soil-chamber system to determine simultaneously δ¹³C_resp and δ¹⁸O_resp. In a Swiss beech forest, a total of 90 chamber measurements with 20 min sampling time each were performed. The instrument measured the δ¹³C and δ¹⁸O of the CO₂ in the chamber headspace at every second with a precision of 0.25‰, resulting in Keeling plots with 1200 data points. In addition, we calculated δ¹³C_resp directly from the flux ratio of ¹³CO₂ and ¹²CO₂. The flux-ratio values were 0.8‰ lower than the Keeling plot intercepts when the flux rates were derived from quadratic curve fits of the CO₂ increase. The δ¹⁸O-Keeling plots showed a significant bending very likely due to the equilibration of chamber CO₂ with the ¹⁸O of surface soil water. Therefore, we used a quadratic curve fit of the Keeling plots to estimate δ¹⁸O_resp. Our results also revealed that δ¹³C_resp was not constant throughout the CO₂ accumulation in the closed soil chambers: there were significant but non-systematic variations in δ¹³C_resp for the first 10 min, and systematic shifts in δ¹³C_resp of on average 1.9 ‰ in the second part of the 20-min measurements. These biases were probably caused by non-steady-state conditions in the soil-chamber system. Our study illustrates that the high temporal resolution of QCLS measurements allows the detection of non-linearities in the isotopic effluxes of CO₂ from the soil due to soil-chamber feedbacks. This information can be used to improve the estimates for δ¹³C_resp and δ¹⁸O_resp.     

C signature of CO2 evolved from incubated maize residues and maize-derived sheep faeces

Analyses of the spatial and temporal variations in the natural abundance of ¹³C are frequently employed to study transformations of plant residues and soil organic matter turnover on sites where long continued vegetation with the C₃-type photosynthesis pathway has been replaced with a C₄-type vegetation (or vice versa). One controversial issue associated with such analyses is the significance of isotopic fractionation during the microbial turnovers of C in complex substrates. To evaluate this issue, C₃-soil and quartz sand were amended with maize residues and with faeces from sheep feed exclusively on maize silage. The samples were incubated at 15 °C for 117 days (maize residues) or 224 days (sheep faeces). CO₂ evolved during incubation was trapped in NaOH and analysed for C isotopic contents. At the end of incubation, 63 and 50% of the maize C was evolved as CO₂ in the soil and sand, respectively, while 32% of the faeces C incubated with soil and with sand was recovered as CO₂. Maize and faeces showed a similar decomposition pattern but maize decomposed twice as fast as faeces. The δ¹³C of faeces was 0.3‰ lower than that of the maize residue (δ¹³C −13.4‰), while the δ¹³C of the C₃-soil used for incubation was −31.6‰. The δ¹³C value of the CO₂ recovered from unamended C₃-soil was similar or slightly lower (up to −1.5‰) than that of the C₃-soil itself except for an initial flush of ¹³C enriched CO₂. The δ¹³C values of the CO₂ from sand-based incubations typically ranged −15‰ to −17‰, i.e. around −3‰ lower than the δ¹³C measured for maize and faeces. Our study clearly demonstrates that the decomposition of complex substrates is associated with isotopic fractionation, causing evolved CO₂ to be depleted in ¹³C relative to substrates. Consequently the microbial products retained in the soil must be enriched in ¹³C.     

Drought alters respired δ13CO2 from autotrophic,but not heterotrophic soil respiration

Many researchers are interested in the variability of root-respired δ¹³CO₂ as an indication of linkages between belowground plant respiration and canopy processes. Most studies in this area have, however, relied upon the assumption that temporal variability of total soil respired δ¹³CO₂ reflects autotrophic soil processes, but in fact few supporting measurements of purely autotrophic soil respiration (partitioned from total soil respiration) are available. Here we use a combination of physical and isotopic partitioning methodologies to track the variability in autotrophic and heterotrophic soil δ¹³CO₂ at five sites in Eastern Canada during a very dry growing season. Three dimensional modeling of soil isotopic transport dynamics in the static sampling chambers allow us to constrain measurement bias and to eliminate non-steady-state effects as a potential driver of observed variability. We provide experimental results that support a pivotal assumption made in prior interpretations of soil δ¹³CO₂ dynamics: we observed minimal isotopic variability in soil microbial δ¹³CO₂ efflux, but appreciable temporal variability in root-respired δ¹³CO₂ at sites where near drought conditions were observed, suggesting that isotopic discrimination is likely linked to seasonal variations in transpirational demand.     

Dual-chamber measurements of δ13C of soil-respired CO2 partitioned using a field-based three end-member model

The contribution of old soil C (SOM) to total soil respiration (R_S) in forest has been a crucial topic in global change research, but remains uncertain. Isotopic methods, such as natural variations in carbon isotope composition (δ¹³C) of soil respiration, are more frequently being applied, and show promise in separating heterotrophic and autotrophic contributions to R_S. However, natural and artificial modification of δ¹³C_Rs can cause isotopic disequilibria in the soil-atmosphere system generating a mismatch between what is usually measured and what process-based models will predict. Here we report the partitioning of the soil surface CO₂ flux in a warm Mediterranean forest into components derived from root, litter/humus, and SOM sources using a new, three end-member mixing model, and compare this with the conventional partitioning into autotrophic and heterotrophic components. The three end-member mixing model takes into account both the discrimination during CO₂ respiration/decomposition of the three components, as well as the fractions of their CO₂ fluxes integrated over the total soil profile mass. In addition, we used a novel dual-chamber technique to ensure that δ¹³C_Rs was subjected to minimal artefacts during measurement.We observed that by using measured soil surface CO₂ concentrations as a baseline level for the dual-chamber operation, it was possible to achieve and monitor the necessary conservation of the soil CO₂ steady-state diffusion conditions during the measurements, without using permanent collars inserted deeply into the soil. When R_S (8.64 g CO₂ m² d⁻¹) was partitioned into two components, the mean autotrophic and heterotrophic respiration was 56 and 44%, respectively. When R_S was partitioned using the three-way model, however, roots, litter/humus, and SOM contributed 30, 33, and 37% of the total flux. Our results confirm that to improve the estimates of the partitioning method, it is important to distinguish the fractional contribution of the long-term SOM-derived flux from younger and more labile sources.     

Three-source partitioning of CO2 efflux from soil planted with maize by C natural abundance fails due to inactive microbial biomass

A theoretical approach to the partitioning of carbon dioxide (CO₂) efflux from soil with a C₃ vegetation history planted with maize (Zea mays), a C₄ plant, into three sources, root respiration (RR), rhizomicrobial respiration (RMR), and microbial soil organic matter (SOM) decomposition (SOMD), was examined. The δ¹³C values of SOM, roots, microbial biomass, and total CO₂ efflux were measured during a 40-day growing period. A three-source isotopic mass balance based on the measured δ¹³C values and on assumptions made in other studies showed that RR, RMR, and SOMD amounted to 91%, 4%, and 5%, respectively. Two assumptions were thoroughly examined in a sensitivity analysis: the absence of ¹³C fractionation and the conformity of δ¹³C of microbial CO₂ and that of microbial biomass. This approach strongly overestimated RR and underestimated RMR and microbial SOMD. CO₂ efflux from unplanted soil was enriched in ¹³C by 2.0‰ compared to microbial biomass. The consideration of this ¹³C fractionation in the mass balance equation changed the proportions of RR and RMR by only 4% and did not affect SOMD. A calculated δ¹³C value of microbial CO₂ by a mass balance equation including active and inactive parts of microbial biomass was used to adjust a hypothetical below-ground CO₂ partitioning to the measured and literature data. The active microbial biomass in the rhizosphere amounted to 37% to achieve an appropriate ratio between RR and RMR compared to measured data. Therefore, the three-source partitioning approach failed due to a low active portion of microbial biomass, which is the main microbial CO₂ source controlling the δ¹³C value of total microbial biomass. Since fumigation-extraction reflects total microbial biomass, its δ¹³C value was unsuitable to predict δ¹³C of released microbial CO₂ after a C₃-C₄ vegetation change. The second adjustment to the CO₂ partitioning results in the literature showed that at least 71% of the active microbial biomass utilizing maize rhizodeposits would be necessary to achieve that proportion between RR and RMR observed by other approaches based on ¹⁴C labelling. The method for partitioning total below-ground CO₂ efflux into three sources using a natural ¹³C labelling technique failed due to the small proportion of active microbial biomass in the rhizosphere. This small active fraction led to a discrepancy between δ¹³C values of microbial biomass and of microbially respired CO₂.     

Uncertainties in static closed chamber measurements of the carbon isotopic ratio of soil-respired CO2

The δ¹³C of soil-respired CO₂ (δ_r) is frequently determined using static closed chamber methods. δ_r is obtained as the intercept of the least squares linear regression of δ vs 1/C*, where measured δ¹³C-CO₂ (δ) and volume fraction of CO₂ (C*) values of chamber headspace samples are used. Theoretically, we show that the variance of the estimate of δ_r can be reduced by extending the 1/C* interval of the regression towards (i) higher or (ii) lower values, or (iii) distributing the 1/C* values optimally within the pre-selected headspace CO₂ sampling time period. Experimental applications of these approaches indicated that: (1) lowering the initial CO₂ level, thereby increasing 1/C*, yielded a positive bias to the δ_r result. (2) It was feasible to obtain lower variance in the δ_r estimate by lowering 1/C* values through extended CO₂ sampling time. We also recommend that each chamber is sampled only once, mainly because this allows freedom to select the sampling times, in order to optimize the distribution of 1/C* values.     

Isotopic discrimination during long-term decomposition in an arid land ecosystem

Discrimination in carbon and nitrogen isotopes of decomposing plant litter in the northern Chihuahuan Desert was determined for a 5-year period. Factors influencing isotopic change were assessed from inter-species comparisons of litter chemistry, mass loss patterns, and isotope values of associated soil. Average δ¹⁵N_litter values of buried roots increased 1.2 and 2.6‰ for Big Blue Stem (Schizachyrium gerardi, grass) and Varital (Drypetes glauca, hardwood) during the study, respectively. Small but inconsistent variations were observed for Slash Pine (Pinus elliotii, conifer) roots. Average δ¹⁵N values of wooden dowels from Ramin (Gonystlylus bancanus, hardwood) increased ca. 2.0‰ during years 1–4, and then decreased slightly during year 5. Changes in δ¹⁵N_litter were independent of N content, and may reflect microbial fractionation or preferential retention of ¹⁵N enriched substrates. Surprisingly, there was no clear relationship between litter N dynamics and C/N ratios. There were no discernable changes in δ¹³C_litter values for Gonystlylus bancanus and Pinus elliotii. Average δ¹³C_litter values for Schizachyrium gerardi decreased ∼2.0‰ during years 0–2 and then increased slightly. In contrast, average δ¹³C_litter values for Drypetes glauca increased ∼0.5‰ from years 0–1 then remained relatively constant until decreasing slightly in year 5. δ¹³C_litter discrimination may have been masked by interfering δ¹³C fractionations or feedbacks between decomposers and litter chemistry. Our data indicate that isotopic discrimination is characteristic of early litter decay stages. These results may highlight aspects of isotope discrimination and nutrient cycling unique to arid land environments. Additional studies will be needed to confirm this.     

Quantifying the contribution of soil organic matter turnover to forest soil respiration, using natural abundance δC

Quantifying the loss of soil carbon through respiration has proved difficult, due to the challenge of measuring the losses associated with the turnover of soil organic matter (SOM) as distinct from autotrophic components. In forest ecosystems the δ¹³C value of respiration from turnover of SOM (δ¹³CR_SOM) is typically 2-4‰ enriched compared with that from roots and associated microbes (δ¹³CR_ROOTS), with that from the litter (δ¹³CR_LITTER) lying between the two. We measured soil respiration at 50 locations in a forest soil and then used differences in isotopic signatures to quantify the proportion of soil respiration arising from the turnover of SOM (fR_SOM) at a subset of 30 locations, chosen randomly. The soil surface CO₂ efflux was collected using an open chamber system supplied with CO₂-free air and the δ¹³C signature (δ¹³CR_S) measured, giving a mean (±SD) value across the site of −26.1 ± 0.58‰. The values of δ¹³CR_ROOTS, δ¹³CR_LITTER and δ¹³CR_SOM were measured at each location by incubation of roots, litter and root-free soil and collection of the CO₂ for isotopic analysis. δ¹³CR_SOM became progressively depleted with length of incubation (1.5‰ after 8 h), so CO₂ was collected after 20 min. The mean value of δ¹³CR_LITTER was −27.2 ± 0.68 ‰, which was indistinguishable from δ¹³CR_ROOTS of −27.6 ± 0.51‰, while δ¹³CR_SOM was −25.1 ± 0.88‰. δ¹³CR_ROOTS and δ¹³CR_SOM measured at each location were used as the end points of a two component mixing model to calculate fR_SOM, giving a mean value for fR_SOM of 0.61 ± 0.28. It was not possible to estimate fR_SOM using the total C contents of the roots and soil which were significantly depleted in ¹³C in comparison to their respired CO₂. However, at seven locations the δ¹³CR_S was slightly enriched compared with δ¹³CR_SOM (mean 0.3‰), which was not considered significantly different so fR_SOM was constrained to 1.0. If these seven rings were excluded mean fR_SOM was 0.49 ± 0.20. We have shown the possibility of using natural abundance ¹³C discrimination to quantify fR_SOM in a forest soil with an input of carbon only from C₃ photosynthesis.     

Characterizing the impact of diffusive and advective soil gas transport on the measurement and interpretation of the isotopic signal of soil respiration

By measuring the isotopic signature of soil respiration, we seek to learn the isotopic composition of the carbon respired in the soil (δ¹³C_R-s) so that we may draw inferences about ecosystem processes. Requisite to this goal is the need to understand how δ¹³C_R-s is affected by both contributions of multiple carbon sources to respiration and fractionation due to soil gas transport. In this study, we measured potential isotopic sources to determine their contributions to δ¹³C_R-s and we performed a series of experiments to investigate the impact of soil gas transport on δ¹³C_R-s estimates. The objectives of these experiments were to: i) compare estimates of δ¹³C_R-s derived from aboveground and belowground techniques, ii) evaluate the roles of diffusion and advection in a forest soil on the estimates of δ¹³C_R-s, and iii) determine the contribution of new and old carbon sources to δ¹³C_R-s for a Douglas-fir stand in the Pacific Northwest during our measurement period. We found a maximum difference of −2.36‰ between estimates of δ¹³C_R-s based on aboveground vs. belowground measurements; the aboveground estimate was enriched relative to the belowground estimate. Soil gas transport during the experiment was primarily by diffusion and the average belowground estimate of δ¹³C_R-s was enriched by 3.8-4.0‰ with respect to the source estimates from steady-state transport models. The affect of natural fluctuations in advective soil gas transport was little to non-existent; however, an advection-diffusion model was more accurate than a model based solely on diffusion in predicting the isotopic samples near the soil surface. Thus, estimates made from belowground gas samples will improve with an increase in samples near the soil surface. We measured a −1‰ difference in δ¹³C_R-s as a result of an experiment where advection was induced, a value which may represent an upper limit in fractionation due to advective gas transport in forest ecosystems. We found that aboveground measurements of δ¹³C_R-s may be particularly susceptible to atmospheric incursion, which may produce estimates that are enriched in ¹³C. The partitioning results attributed 69-98% of soil respiration to a source with a highly depleted isotopic signature similar to that of water-soluble carbon from foliage measured at our site.     

Dynamics of C natural abundance in wood decomposing fungi and their ecophysiological implications

Factors that affect the δ¹³C values of fungi need to be analyzed for the progress of isotope-based studies of food-chain or organic matter dynamics in soils. To analyze the factors that control δ¹³C values of the fungal body, basidiomycete and ascomycete species were grown on a beechwood substrate (six species) and in glucose medium (nine species), and the δ¹³C value of produced fungal body was compared to that of the carbon source. The ¹³C enrichment (Δδ¹³C) in the fungal aggregates compared to the decomposed wood varied from 1.2 to 6.3‰ among six species. In the glucose substrate experiment, the degree of ¹³C enrichment in the hyphal mat was relatively small and varied from −0.1 to 2.8‰ among nine basidiomycetes species depending on their growth stage. Calculated δ¹³C values of the respired CO₂ were lower than those of the hyphal mat, organic metabolites and the glucose used. The degree of ¹³C enrichment was affected by fungal species, substrate and growth stage. Fungal internal metabolic processes are the plausible mechanism for the observed isotopic discrimination between fungal bodies and substrates. Especially, dark fixation of ambient CO₂ and kinetic isotope fractionation during assimilation and dissimilation reactions could well explain Δδ¹³C dynamics in our experiments. Through the analysis of field Δδ¹³C, we could know undisturbed fungal status about starvation, aeration and type of decomposition.     

The stable carbon isotope composition of soil organic carbon and pedogenic carbonates along a bioclimatic gradient in the Palouse region,Washington State,USA

Isotopic signatures of soil components are commonly used to infer past ecologic and climatic shifts in the soil record. The theory behind the fractionation of isotopes that occurs during ecosystem processes is well understood; however, few isotopic studies have explored ecosystem relationships in modern soils. We discuss relationships of stable carbon isotopic signatures in plant tissue, soil organic carbon (SOC), laboratory-respired CO₂, and modern carbonates at 10 sites (seven containing pedogenic carbonates) along a C₃-dominated climatic gradient (mean annual precipitation (MAP) ranging from 200 to 1000 mm) in the Palouse region of eastern Washington state.A horizon soil organic carbon (SOC) δ¹³C values varied from −24.3‰ to −25.9‰ PDB. Values in the arid portion of the gradient (200 to approximately 500 mm MAP) generally decreased and linear regression of SOM ¹³C vs. MAP was significant (r²=0.71, p=0.02). Trends in plant-¹³C of two grass species (Agropyron spicatum and Festuca idahoensis) found throughout this portion of the gradient were similar to that of SOC. Mean pedogenic carbonate δ¹³C values varied from −4.1‰ to −10.8‰ PDB. Linear regression was significant for carbonate ¹³C vs. MAP (r²=0.79, p=0.007), estimated above-ground productivity (r²=0.88, p=0.002) and soil carbon content (r²=0.83, p=0.004). Carbonate δ¹³C values at the most arid site exhibited higher variability than other sites (presumably due to greater spatial variation in plant respiration vs. atmospheric diffusion). Our data suggest that carbon isotopic relationships among ecosystem components may prove useful in determining ecosystem level properties in modern systems, and potentially in ancient systems as well.     

Nitrogen fixation by biological soil crusts and heterotrophic bacteria in an intact Mojave Desert ecosystem with elevated CO2 and added soil carbon

Fixation of N by biological soil crusts and free-living heterotrophic soil microbes provides a significant proportion of ecosystem N in arid lands. To gain a better understanding of how elevated CO₂ may affect N₂-fixation in aridland ecosystems, we measured C₂H₂ reduction as a proxy for nitrogenase activity in biological soil crusts for 2 yr, and in soils either with or without dextrose-C additions for 1 yr, in an intact Mojave Desert ecosystem exposed to elevated CO₂. We also measured crust and soil δ¹⁵N and total N to assess changes in N sources, and δ¹³C of crusts to determine a functional shift in crust species, with elevated CO₂. The mean rate of C₂H₂ reduction by biological soil crusts was 76.9±5.6 μmol C₂H₄ m⁻² h⁻¹. There was no significant CO₂ effect, but crusts from plant interspaces showed high variability in nitrogenase activity with elevated CO₂. Additions of dextrose-C had a positive effect on rates of C₂H₂ reduction in soil. There was no elevated CO₂ effect on soil nitrogenase activity. Plant cover affected soil response to C addition, with the largest response in plant interspaces. The mean rate of C₂H₂ reduction in soils either with or without C additions were 8.5±3.6 μmol C₂H₄ m⁻² h⁻¹ and 4.8±2.1 μmol m⁻² h⁻¹, respectively. Crust and soil δ¹⁵N and δ¹³C values were not affected by CO₂ treatment, but did show an effect of cover type. Crust and soil samples in plant interspaces had the lowest values for both measurements. Analysis of soil and crust [N] and δ¹⁵N data with the Rayleigh distillation model suggests that any plant community changes with elevated CO₂ and concomitant changes in litter composition likely will overwhelm any physiological changes in N₂-fixation.     

Changes in the carbon and nitrogen isotopic composition of organic matter in soils of different thermal stability after free-air CO2 enrichment for three years

The hypothesis that the biological availability of soil organic matter (SOM) pools is inversely proportional to their thermal stability was tested using the isotopic difference between the atmospheric CO₂ (δ¹³C = ?8.0‰) and ¹³C-enriched CO₂ (δ¹³C = ?47‰) fertilizers, as well as ¹⁵N-labeled fertilizers. The soil samples from spring wheat plots subjected to treatment with ambient (370 ppm) and elevated (540 ppm) CO₂ concentrations for three years were analyzed by the thermogravimetric method. Based on the weight loss, five SOM pools were distinguished where the total C and N contents and isotopic compositions (δ¹³C and (δ¹⁵N) were determined. The contents of new C and N and their mean residence times in pools were calculated. The incorporation of ¹³C and ¹⁵N and their turnover rates did not depend on the thermal stability of the SOM pools, which disproved the hypothesis being tested.     

Three‐source partitioning of CO2 efflux from maize field soil by 13C natural abundance

A natural‐¹³C‐labeling approach—formerly observed under controlled conditions—was tested in the field to partition total soil CO₂ efflux into root respiration, rhizomicrobial respiration, and soil organic matter (SOM) decomposition. Different results were expected in the field due to different climate, site, and microbial properties in contrast to the laboratory. Within this isotopic method, maize was planted on soil with C₃‐vegetation history and the total CO₂ efflux from soil was subdivided by isotopic mass balance. The C₄‐derived C in soil microbial biomass was also determined. Additionally, in a root‐exclusion approach, root‐ and SOM‐derived CO₂ were determined by the total CO₂ effluxes from maize (Zea mays L.) and bare‐fallow plots. In both approaches, maize‐derived CO₂ contributed 22% to 35% to the total CO₂ efflux during the growth period, which was comparable to other field studies. In our laboratory study, this CO₂ fraction was tripled due to different climate, soil, and sampling conditions. In the natural‐¹³C‐labeling approach, rhizomicrobial respiration was low compared to other studies, which was related to a low amount of C₄‐derived microbial biomass. At the end of the growth period, however, 64% root respiration and 36% rhizomicrobial respiration in relation to total root‐derived CO₂ were calculated when considering high isotopic fractionations between SOM, microbial biomass, and CO₂. This relationship was closer to the 50% : 50% partitioning described in the literature than without fractionation (23% root respiration, 77% rhizomicrobial respiration). Fractionation processes of ¹³C must be taken into account when calculating CO₂ partitioning in soil. Both methods—natural ¹³C labeling and root exclusion—showed the same partitioning results when ¹³C isotopic fractionation during microbial respiration was considered and may therefore be used to separate plant‐ and SOM‐derived CO₂ sources.     

Soil CO<Subscript>2</Subscript> sources above a subterranean cave—Pisani rov (Postojna Cave,Slovenia)

Purpose

The objective of this research is to detect abiotic sources of soil CO₂ above a subterranean cave in the Slovenian karst region.

Materials and methods

The research was performed in the forest above Pisani rov (Postojna Cave) near the town of Postojna (SW Slovenia) and also in the cave. Soil gas, atmospheric air and cave air carbon stable isotope composition (δ¹³C_CO2) and CO₂ concentration were measured. Sampling and measurements were performed bi-monthly at the test and control sites above the cave. The abiotic source of soil CO₂ was estimated using a stable isotope mass balance calculation.

Results and discussion

Similar seasonal patterns of soil CO₂ and δ¹³C_CO2 values were observed at both the test and control sites until spring, with higher levels of CO₂ observed in summer and lower in winter. The δ¹³C_CO2 showed the opposite trend, i.e. lower values (?26 to ?20 ‰) in summer and higher values (up to ?17 ‰) in winter and early spring. In spring, the soil CO₂ concentration decreases and the δ¹³C_CO2 value increases only at the control site. A time series of a modelled “isotopically light” endmember revealed large shifts in the data values, due to the presence of an abiotic CO₂ source. Results suggest that the subterranean CO₂ pool and its ventilation is the main source of soil CO₂, accounting for up to 80 % of the soil gas during cold periods.

Conclusions

Ventilation from subterranean cavities is an important source of soil CO₂ in karstic areas and should be taken into account during carbon cycling studies.

     

Evaluating N/N and C/C isotope ratio analysis to investigate trophic relationships of elaterid larvae (Coleoptera: Elateridae)

Carbon and nitrogen isotope ratios in consumer tissues can be used to analyse the diet and trophic level of soil animals. However, life history traits may significantly influence stable isotope patterns. We evaluated in a series of experiments how stable isotope ratios of carbon (¹³C/¹²C) and nitrogen (¹⁵N/¹⁴N) at natural abundance can be used to study the diet and trophic position of long-lived macro-invertebrates, elaterid larvae, which are major below-ground herbivores. Small, but significant differences in δ¹³C signatures were found between the larvaes’ anterior and posterior body segments, whereas exuvia reflected the body's overall isotopic composition. The species-specific trophic shift (±SE) in δ¹⁵N for Agriotes obscurus and Agriotes sputator (1.62±0.24‰ and 1.08±0.27‰, respectively) was significantly lower than “mean enrichment estimates” reported in the literature, showing the limited applicability of such generalised estimates in studies of invertebrate trophic ecology. To avoid false-positive assignments to two trophic levels due to variation in δ¹⁵N values, a minimum sample size of three and five individuals for A. obscurus and A. sputator, respectively, was needed to reduce this risk to below α=5%. Keeping elaterid larvae for up to 128 days without food did not affect their isotopic signatures, in contrast to previous studies on starving animals. Switching wireworms to isotopically different diets induced changes in their isotopic signatures within 2 weeks. Changes, however, were significant only when the isotopic difference between diets was large. We conclude that experimental studies evaluating how specific life history traits affect stable isotope signatures in consumers have to precede any interpretation of stable isotope data gathered in the field.     

Carbon stable isotope fractionation and trophic transfer of fatty acids in fungal based soil food chains

Stable isotope analysis has been used as a powerful tool in food web studies in terrestrial ecosystems. In addition the occurrence and abundance of fatty acids may serve as indicator for feeding strategies of soil animals. Here we combine both approaches and investigate the fatty acid composition, δ¹³C values of bulk tissues and individual fatty acids in soil organisms. The fungi Chaetomium globosum and Cladosporium cladosporioides were isotopically labelled by fructose derived from either C₃ or C₄ plants, and the fungal-feeding nematode Aphelenchoides sp. was reared on C. globosum. Fungi and nematodes were used as diet for the Collembolan Protaphorura fimata. The sugar source was fractionated differently by fungal lipid metabolism in a species-specific manner that points to a sensitivity of physiological processing to the non-random distribution of ¹³C/¹²C isotopes in the molecule. As a general trend stearic acid (18:0) was depleted in ¹³C compared to the precursor palmitic acid (16:0), whereas its desaturation to oleic acid (18:1 ω9) favoured the ¹³C-rich substrate.Fatty acid profiles of P. fimata varied due to food source, indicating incorporation of dietary fatty acids into Collembolan tissue. Individuals feeding on fungi had lower amounts in C20 fatty acids, with monoenoic C20 forms not present. This pattern likely separates primary consumers (fungivores) from predators (nematode feeders). The isotopic discrimination in ¹³C for bulk Collembola ranged between −2.6 and 1.4‰ and was dependent on fungal species and C₃/C₄ system, suggesting differences at metabolic branch points and/or isotope discrimination of enzymes. Comparison of δ¹³C values in individual fatty acids between consumer and diet generally showed depletion (i.e. de novo synthesis) or no changes (i.e. dietary routing), but the fractionation was not uniform and affected by the type of ingested food. Fatty acid carbon isotopes were more variable than those of bulk tissues, likely due to both the distrimination by enzymes and the different lipid origin (i.e. neutral or polar fraction).     

Effluxed CO2-C from sterilized and unsterilized treatments of a calcareous soil

Soil inorganic carbon (C) represents a substantial C pool in arid ecosystems, yet little data exist on the contribution of this pool to ecosystem C fluxes. A closed jar incubation study was carried out to test the hypothesis that CO₂-¹³C production and response to sterilization would differ in a calcareous (Mojave Desert) soil and a non-calcareous (Oklahoma Prairie) soil due to contributions of carbonate-derived CO₂. In addition to non-sterilized controls, soils were subjected to sterilization treatments (unbuffered HgCl₂ addition for Oklahoma soil and unbuffered HgCl₂ addition, buffered HgCl₂ addition, and autoclaving for Mojave Desert soil) to decrease biotic respiration and more readily measure abiotic CO₂ flux. Temperature and moisture treatments were also included with sterilization treatments in a factorial design.The rate of CO₂ production in both soils was significantly decreased (36-87%) by sterilization, but sterilization treatments differed in effectiveness. Sterilization had no significant effect on effluxed CO₂-¹³C values in the non-calcareous Oklahoma Prairie soil and autoclaved Mojave Desert soil as compared to their respective non-sterilized controls. However, sterilization significantly altered CO₂-¹³C values in Mojave Desert soil HgCl₂ sterilization treatments (both buffered and non-buffered). Plots of 1/CO₂ versus CO₂-δ¹³C (similar to Keeling plots) indicated that the source CO₂-δ¹³C value of the Oklahoma Prairie soil treatments was similar to the δ¹³C value of soil organic matter [(SOM); −17.76‰ VPDB] whereas the source for the (acidic) unbuffered-HgCl₂ sterilized Mojave Desert soil was similar to the δ¹³C value of carbonates (−0.93‰ VPDB). The source CO₂-δ¹³C value of non-sterilized and autoclaved (−18.4‰ VPDB) Mojave Desert soil treatments was intermediate between SOM (−21.43‰ VPDB) and carbonates and indicates up to 13% of total C efflux may be from abiotic sources in calcareous soils.     

Continuous flow isotope ratio mass spectrometry of carbon dioxide trapped as strontium carbonate

Abstract

The isotopic signal provided by differential discrimination against atmospheric carbon dioxide (¹³CO₂) by C₃ and C₄ plant photosynthetic pathways is being widely used to study the processes of carbon (C) fixation, soil organic matter formation, and mineralization in nature. These studies have been facilitated by the availability of automated C and nitrogen (N) combustion analyzers (ANCA) combined with continuous flow isotope ratio mass spectrometers (CFIRMS). Analysis of ¹³CO₂ in these instruments requires consistent sample mass for best precision, a requirement that is easily satisfied for soil and tissue samples by adjusting sample weight. Consistent CO₂ sample size is much more difficult to achieve using gas handling systems for samples of headspace gases when CO₂ concentrations vary widely. Long storage of gaseous samples also is difficult. Extended respiration studies are most easily conducted by trapping CO₂ in alkali and conversion to an insoluble carbonate. Thermal decomposition of the carbonate in an on‐line ANCA allows consistent and optimal CO₂ sample mass to be obtained. The use of precipitated carbonates also facilitates storage of samples and enables full automation of sample analysis using an ANCA interfaced to a CFIRMS. Calcium (Ca), strontium (Sr), and barium (Ba) carbonates were tested. Strontium carbonate (SrCO₃) with the addition of vanadium pentoxide (V₂O₅) as a combustion catalyst was found most suitable.     

The effect of soil moisture,soil particle size,litter layer and carbonic anhydrase on the oxygen isotopic composition of soil‐released CO2

Soil respiration and photosynthesis are the two largest carbon dioxide (CO₂) fluxes between terrestrial ecosystems and the atmosphere and, therefore, the dominant processes influencing the oxygen isotopic composition of atmospheric CO₂. The characterization of temporal and spatial variations of plant and soil‐related fluxes of different oxygen isotopologues of CO₂ (¹²C¹⁶O₂; ¹²C¹⁶O¹⁸O) is relevant to constraining the global carbon budget. The oxygen isotopic composition of soil‐respired CO₂ is controlled by its release rate, the degree of isotopic equilibrium with soil water and the diffusional transport of CO₂. The hypothesis of this study was that, as well as soil moisture, the soil particle size, the presence of an organic litter layer and the enzyme carbonic anhydrase (CA) would have a significant impact on the oxygen isotopic composition of soil‐released CO₂. We tested this hypothesis with soil microcosm experiments on columns of medium and fine sand. Soil water content and soil texture influenced the isotopic composition of soil‐released CO₂ significantly. A litter layer had a significant effect on the isotopic composition of water vapour but not on CO₂ released from soil. In the absence of CA, oxygen isotope equilibration between the CO₂ invasion flux and soil water was insignificant, whereas in the presence of CA about 55% of the CO₂ invading the soil exchanged oxygen isotopes with soil water. Our findings highlight the importance of small‐scale variability of soil attributes for the oxygen isotopic composition of soil‐released CO₂ as well as the strong impact of CA activity in soils.